GPD2 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the GPD2 gene in the NCI-H1975 non-small cell lung cancer (NSCLC) cell line. This product provides a genetically heterogeneous pool of edited cells, each carrying CRISPR/Cas9-mediated gene disruptions, enabling loss-of-function studies of mitochondrial glycerol-3-phosphate dehydrogenase without selection for a single clonal genotype. The polyclonal format preserves population-level diversity and is suitable for experiments where bulk knockout effects are assayed.
The NCI-H1975 host cell line is a well-characterized human lung adenocarcinoma epithelial model established from a female NSCLC patient. It harbors activating EGFR L858R and T790M mutations, conferring constitutive kinase activity and resistance to first-generation tyrosine kinase inhibitors. These genetic features make NCI-H1975 a critical tool for investigating oncogenic signaling, drug resistance mechanisms, and metabolic adaptations in lung cancer. The epithelial origin and adenocarcinoma histology further contextualize its use in tumor biology and therapeutic development.
The GPD2 gene encodes mitochondrial glycerol-3-phosphate dehydrogenase, a flavin adenine dinucleotide (FAD)-dependent enzyme that catalyzes the oxidation of glycerol-3-phosphate to dihydroxyacetone phosphate. This reaction is coupled to the reduction of FAD to FADH2, which donates electrons directly to coenzyme Q of the mitochondrial respiratory chain, thereby linking glycolysis-derived glycerol-3-phosphate to oxidative phosphorylation. GPD2 function is integral to the glycerol phosphate shuttle, working in concert with cytosolic GPD1 to maintain cellular redox balance and ATP production. The enzyme is transcriptionally regulated by PPAR??, PPAR??, PGC-1??, and HIF-1??, and its activity is modulated by insulin signaling. Downstream, GPD2 influences the NAD+/NADH ratio, drives ATP synthesis, and modulates reactive oxygen species (ROS) generation through its impact on electron transport chain flux.
In the NCI-H1975 background, GPD2 knockout disrupts the glycerol phosphate shuttle, forcing metabolic rerouting and potentially compromising mitochondrial respiration. Given the reliance of NSCLC cells on metabolic plasticity for growth and survival, loss of GPD2 may reveal vulnerabilities in energy metabolism and redox homeostasis. This model allows researchers to dissect the intersection of EGFR-driven signaling and mitochondrial function, providing insights into how oncogenic mutations shape metabolic dependencies. The EGFR T790M mutation, in particular, is associated with altered metabolic profiles, making this knockout system valuable for studying resistance-associated metabolic shifts.
This polyclonal knockout cell product is suitable for a broad range of applications in cancer metabolism and mitochondrial research. Representative assays include Seahorse metabolic flux analysis to assess oxygen consumption and extracellular acidification, glycerol-3-phosphate dehydrogenase activity measurements, and ATP bioluminescence quantification. ROS detection and cell proliferation/viability assays can define the functional consequences of GPD2 loss. Expression analysis by RT-qPCR and western blotting can monitor metabolic markers such as HIF-1??, PPAR??, and PGC-1??. These cells are especially useful for drug sensitivity screening, oxidative stress studies, and investigations into metabolic reprogramming in NSCLC. For additional information, please contact Ascent Research.
